健心颗粒对气虚血瘀型心力衰竭大鼠心肌重构及CaN/NFATc3通路的影响

Effects of Jianxin Granule on Myocardial Remodeling and CaN/NFATc3 Pathway in Rats with Heart Failure of Qi Deficiency and Blood Stasis

目的 观察健心颗粒对气虚血瘀型心力衰竭(心衰)大鼠心肌重构的影响,并探讨其对钙调磷酸酶(Ca N)/活化T细胞核因子c3(NFATc3)通路的影响。方法 将24只雄性SD大鼠随机分为假手术组(Sham组)、模型组和干预组,每组8只。模型组和干预组结扎冠脉左前降支后,配合饥饿和力竭游泳的方式构建气虚血瘀型心衰模型,Sham组大鼠冠脉左前降支仅穿线不接扎。干预组予以健心颗粒3.2 g/(kg·d),Sham组和模型组均予以2 mL生理盐水灌胃,每日1次。干预4周后,行右颈动脉插管测定大鼠血流动力学参数[左室收缩末压(LVSP)、左室舒张末压(LVEDP)、左室压力最大上升速率(+dP/dtmax)及左室压力最大下降速率(-dP/dtmax)],测量心脏重量,计算心脏重量指数(HMI),心肌组织行Masson、HE染色分别测定心肌梗死面积百分比、心肌细胞横截面积。留取大鼠心肌组织行CaN的活性检测,免疫组化和Western Blot测定CaNA亚基(CaNA)、CaNB亚基(CaNB)、胞核和胞浆NFATc3及β肌球蛋白重链(β-MHC)、B型利钠肽(BNP)的表达。结果 Sham组大鼠一般情况良好,心肌细胞形态轮廓清晰,排列整齐,少许的炎细胞浸润;模型组大鼠疲乏、萎靡,活动度少,呼吸急促,心肌组织受损严重,心肌细胞轮廓模糊,呈现无序化排列,大量炎症细胞浸润于心肌组织间;与模型组比较,干预组精神状态改善,静息状态下无呼吸急促,活动度增加,心肌形态较模型组完整,排列相对有序,细胞间炎症细胞浸润减少。与Sham组大鼠比较,模型组大鼠LVSP、+dP/dtmax、-dP/dtmax降低,LVEDP、心肌梗死面积百分比、横截面积、HMI、心肌组织CaN活性、β-MHC、BNP蛋白表达、胞核NFATc3蛋白表达以及胞核NFATc3/胞浆NFATc3升高(均P<0.05)。与模型组比较,干预组大鼠LVSP、+dP/dtmax、-dP/dtmax升高,LVEDP、心肌梗死面积百分比、横截面积、HMI、心肌组织CaN活性、β-MHC、BNP蛋白表达、胞核NFATc3蛋白表达以及胞核NFATc3/胞浆NFATc3降低(均P<0.05)结论 健心颗粒可改善气虚血瘀型心衰大鼠的心功能,改善心肌重构,下调心肌β-MHC、BNP的表达,其机制可能与抑制心肌组织CaN的活性,进而下调NFATc3的核转位有关。

Objective To observe the effects of Jianxin Granule on myocardial remodeling in rats with heart failure of qi deficiency and blood stasis,and to explore the mechanism of Jianxin Granule on calcineurin(CaN)/nuclear factor c3 of activated T cells(NFATc3)pathway.Methods Twenty-four male SD rats were randomly divided into Sham group,model group and intervention group,8 rats in each group.The rats in the model group and intervention group were undergone ligation of the left anterior descending coronary artery,and the heart failure model of qi deficiency and blood stasis was established by starvation and exhaustive swimming.The left anterior descending coronary artery was only threaded without ligation in the Sham group.The rats in intervention group were given Jianxin Granule 3.2 g·kg-1·d-1 by gastrogavage once a day,and rats in Sham group and model group were given 2 mL normal saline by gastrogavage once a day.After 4 weeks intervention,rats hemodynamic parameters including left ventricular peak systolic pressure(LVSP),left ventricular end-diastolic pressure(LVEDP),maximal rate of the increase of left ventricular pressure(+dP/dtmax)and maximal rate of the decrease of left ventricular pressure(-dP/dtmax)were detected by cannulated though the right carotid artery,the heart weight was measured,the heart mass index(HMI)was calculated,and the myocardial tissue was stained by Masson and HE to measure infarct size and myocardial cross-sectional area respectively.Myocardial tissue was collected to detect the activity of CaN.Immunohistochemistry and Western Blot were used to detect calcineurin A subunit(CaNA),calcineurin B subunit(CaNB),nuclear and cytosol NFATc3,β-myosion heavy chain(β-MHC)and B-type natriuretic peptide(BNP).Results The rats in Sham group were generally in good condition,with clear outline and neat arrangement of cardiomyocytes,and a little inflammatory cell infiltration.The rats in the model group were fatigue,lethargy,low mobility,and shortness of breath,the myocardium was severely damaged,the outline of myocardium was blurred and disordered,and a large number of inflammatory cells infiltrated the myocardium.Compared with the model group,the mental state of the intervention group was improved,there was no shortness of breath at rest,the activity was increased,the myocardial morphology was more complete than the model group,the arrangement was relatively orderly,and the infiltration of intercellular inflammatory cells was reduced.Compared with the Sham group,LVSP,+dP/dtmax,-dP/dtmax decreased,LVEDP,myocardial infarct size and cross-sectional area,HMI,the activity of CaN in the myocardial tissue,the expression ofβ-MHC and BNP,the expression of nuclear NFATc3 and nuclear NFATc3/cytosol NFATc3 increased in the model group(P<0.05).Compared with the model group,LVSP,+dP/dtmax,-dP/dtmax increased,LVEDP,myocardial infarct size and cross-sectional area,HMI,the activity of CaN in the myocardial tissue,the expression ofβ-MHC and BNP,the expression of nuclear NFATc3 and nuclear NFATc3/cytosol NFATc3 decreased in the intervention group(P<0.05).Conclusions Jianxin Granule could improve cardiac function and myocardial remodeling,down-regulating the expression ofβ-MHC and BNP in heart failure rats with qi deficiency and blood stasis.Its mechanism might be related to inhibiting the activity of CaN in myocardial tissue and down-regulating the nuclear translocation of NFATc3.